Identification of Rho(D) antigen in polyacrylamide gels by an enzyme-linked immunoassay.
Human erythrocyte membranes were solubilized in sodium dodecyl sulfate at 100 degree C and subjected to polyacrylamide gel electrophoresis. The gels were sliced into segments and each segment was incubated with anti-Rho(D) IgG, washed, and then incubated with goat anti-human IgG covalently linked to alkaline phosphatase. Para-nitrophenyl phosphate was added to each slice and the absorbance of the solution surrounding each slice was measured at 405 nm. This technique demonstrated that the Rho (D) antigen is a protein with a mol, wt between 13, 000 and 30,000. This method should be applicable to the investigation of other membrane-bound antigens.
D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D004910:Erythrocyte Membrane / Q000276:immunology; D006801:Humans; D008970:Molecular Weight; D012204:Rh-Hr Blood-Group System / Q000276:immunology*
D012204:Rh-Hr Blood-Group System; C026410:Rho(D) antigen